微型dna条形码用于猛禽物种鉴定.docx

微型DNA条形码用于猛禽物种鉴定 高航 摘要：以猛禽为研究对象，首先采用模拟实验 (in silico an alysis)，将BOLD数据库中230个样本的序 列拆成 3或6段相同大小(216 bp或 108 bp)的片段，选用 Full-barcode、Mini-barcode216-1 和 Mini-barcode 108-1三个片段数据集以K2P为参数构建NJ树，并进行遗传距离分析得出解析度的准确性依次为 84.1%、78.3%、75.2% ;然后进行实验验证 (Empirical test)，设计PCF引物扩增40个样本的DNA条形 码指定序列5端114 bp、中间103 bp和3 端121 bp,分别以单一片段和两两组合后的片段序列数据集 为分析基础计算物种鉴定的准确性，得出 114+103 bp合并序列为85.0%、114+121 bp为85.7%。因此， 微型DNA条形码(Minimalist barcod e)鉴定猛禽物种是可行的，和 DNA条形码一样均可作为猛禽物种 鉴定的可靠依据。 关键词：猛禽；DNA条形码；微型条形码；物种鉴定 Minimalist-barcoding on the SpeciesIdentification of Raptors Abstract This study testified the feasibility of species identification of Raptors with minimalist barcodes. We first tested the in silico performance of ‘-bmairnciodes (216 bp or 108 bp) in specie-lsevel identifications of 230 COI sequences from BOLD system, and calculated the resolution of three segments of full-barcode, mini-barcode 216-1 and mini-barcode 108-1 by neighbour-joining (NJ) analysis. With the segments decreasing, the resolution of species identification lowered (84.1%, 78.3% and 75.2%, respectively). However, the choice of length and position of mini-barcodes was important in their ability to discriminate among species. We gen erated 111 min i-barcodes (?100 bp) from 5 end, middle and 3 end of DNA barcode, respectively. The combinated segments of 114+103 bp and 114+121 bp showed higher accuracy at 85.0% and 85.7%, as compared to others in species-level identification by NJ analysis. The mini-barcodes which produced species-level resolution congruent with DNA barcode were also valuable for the identification of the raptors species whose DNA were degraded. Key words:Raptors; DNA barcode; Minimalist-barcode; Species identification 猛禽是鸟类六大生态类群之一， 涵盖了鸟类传统分类系统中隼形目和鸮形目的所有种， 包括鹗、 （ 李湘涛， （ 李湘涛， 2004）。随着 年龄、性别、季节和亚种的不同，猛禽在形态上会出现显著变异，对这些猛禽的准确识别鉴定便成 为了生物保护工作的基础。过去，人们一直采用传统的形态学分类方法，该方法的一些缺陷造成了 生物保护工作的局限 （莫帮辉等， 2008）。随着现代分子生物技术的发展，利用 DNA 序列片断信息对 生物进行种类鉴定、物种分类地位及种间亲缘关系探讨正在成为一个主流手段。其中， Hebert提出 的线粒体COI基因指定片段（648 bp）为基础的DNA条形码（DN